RESUMO
1. In the present study we provide evidence for a saturable, Mg2+/ATP- and temperature-dependent, tetrabenazine-, dopamine-, and amphetamine-sensitive uptake of 1-methyl-4-phenylpyridinium ion (MPP+) in synaptic vesicles from mouse striatum. 2. Similarity in the properties of the vesicular uptake suggests that in the striatum dopamine and MPP+ share the vesicular carrier. 3. The presence of MPP+ vesicular uptake in dopamine-rich regions such as striatum, olfactory, tubercles and hypothalamus, as well as its absence in cerebellum, cortex and pons-medulla, suggest that monoamine vesicular carriers differ between highly and poorly dopamine-innervated regions. 4. The restriction of active MPP+ uptake to the dopaminergic regions, which reflects the previously shown distribution of [3H]-MPP+ binding sites in mouse brain membranes, indicates MPP+ as a marker of the vesicular carrier for dopamine in dopaminergic neurones. 5. A role in MPP+ neurotoxicity is suggested for this region-specific, vesicular storage of the toxin.
Assuntos
1-Metil-4-Fenil-1,2,3,6-Tetra-Hidropiridina/análogos & derivados , Dopaminérgicos/toxicidade , Dopamina/fisiologia , Doenças do Sistema Nervoso/induzido quimicamente , Compostos de Piridínio/metabolismo , Vesículas Sinápticas/metabolismo , Anfetamina/farmacologia , Animais , Biomarcadores , Encéfalo/efeitos dos fármacos , Encéfalo/metabolismo , Corpo Estriado/efeitos dos fármacos , Corpo Estriado/metabolismo , Dopamina/farmacologia , Técnicas In Vitro , Intoxicação por MPTP , Masculino , Camundongos , Microscopia Eletrônica , Doenças do Sistema Nervoso/metabolismo , Tetrabenazina/farmacologiaRESUMO
A recent study reported a possible association between allele 1 of the dopamine D3 receptor gene and bipolar affective disorder using the haplotype relative risk approach. In attempt to replicate these findings, we used similar family-based methods, such as the Haplotype-Based Haplotype Relative Risk method and the Transmission Disequilibrium Test, in a sample of 44 bipolar probands from Sardinia with both parents available. Using the Bal I restriction enzyme site polymorphism of Lannfelt et al. (1992), no differences were found between transmitted and non-transmitted alleles and no evidence of linkage disequilibrium was observed.
Assuntos
Transtorno Bipolar/genética , Receptores de Dopamina D2/genética , Alelos , Genótipo , Haplótipos , Heterozigoto , Humanos , Desequilíbrio de Ligação , Polimorfismo Genético , Receptores de Dopamina D3RESUMO
We aimed to ascertain whether pyrethroid insecticides could influence the vesicular transport of the excitatory amino acid glutamate. The incubation of rat cortical synaptic vesicles with resmethrin and permethrin, consistently stimulated both ATP-dependent and -independent uptake of [3H]glutamate, while not evoking depletion of its vesicular content. Both processes were counteracted by valinomycin, a dissipator of the transmembrane potential gradient (deltapsi(sv)). Meanwhile, the vesicular influx of 36Cl- anions was impaired by pyrethroid concentrations which did not affect the ATP-dependent uptake of [14C]methylamine, as a marker for the proton gradient (deltapH). Thus, the stimulation of glutamate transport appeared to involve mainly the deltapsi(sv). A self-attenuating effect of selected pyrethroids on putatively enhanced excitatory transmission in severe intoxication is suggested.
Assuntos
Ácido Glutâmico/farmacocinética , Piretrinas/metabolismo , Vesículas Sinápticas/efeitos dos fármacos , Vesículas Sinápticas/metabolismo , Trifosfato de Adenosina/metabolismo , Animais , Transporte Biológico/efeitos dos fármacos , Radioisótopos de Carbono , Cloretos/farmacocinética , Inseticidas/farmacologia , Ionóforos/farmacologia , Masculino , Metilaminas/farmacocinética , Permetrina , Piretrinas/farmacologia , Ratos , Ratos Sprague-Dawley , Trítio , Valinomicina/farmacologiaRESUMO
[3H] N-Methyl-4-phenylpyridinium ion (MPP+) binds with a fully reversible, high affinity process to a population of sites mainly localized in the mouse striatum (Bmax = 168 +/- 15 fmol/mg protein, KD = 1.4 +/- 0.4 nM). The majority of specifically-bound radioactivity was localized in the synaptosomal fraction. Unilateral, striatal denervation with 6-hydroxydopamine (6-OHDA) markedly (by 65-70%) decreased the number of [3H]MPP+ sites. Besides dopamine, the vesicular markers tyramine, tetrabenazine and reserpine inhibited [3H]MPP+, while mazindol was a poor displacer. Adenosine triphosphate (ATP) and Mg(2+)-ions did not affect [3H]MPP+ binding. It is concluded that these sites may represent a marker of striatal storage vesicles for dopamine.
Assuntos
1-Metil-4-fenilpiridínio/metabolismo , Corpo Estriado/metabolismo , Sinaptossomos/metabolismo , Animais , Sítios de Ligação , Ligação Competitiva , Membrana Celular/metabolismo , Cinética , Masculino , Camundongos , Especificidade de Órgãos , Frações Subcelulares/metabolismo , TrítioRESUMO
Low (up to 100 nM) and high (approximately 100 microM) concentrations of lanthanides and Ca2+-ions, respectively, stimulated [3H]tyramine binding ([3H]TY) to rat striatal membranes, a putative marker for the vesicular transporter of dopamine. On the other hand, lanthanides (approximately 100 microM) inhibited or stimulated TY binding in striatal and extrastriatal (cortex, cerebellum) tissues, respectively. The binding increases by lanthanum (La3+) appeared to depend on endogenous Ca2+, whereas, those induced in EDTA-pretreated membranes were Ca2+-independent. The La3+-induced, apparent increase in the Bmax for [3H]TY binding seemed to reflect a retarded rate of dissociation of the ligand from its targets, rather than a larger availability of functionally-relevant, vesicular transport-related TY sites. This indicates uncertain mechanisms of present La3+ effects.
Assuntos
Adrenérgicos/farmacologia , Corpo Estriado/química , Corpo Estriado/metabolismo , Metais Terras Raras/farmacologia , Tiramina/farmacologia , Trifosfato de Adenosina/farmacologia , Adrenérgicos/metabolismo , Animais , Cálcio/metabolismo , Cálcio/farmacologia , Cerebelo/química , Cerebelo/efeitos dos fármacos , Cerebelo/metabolismo , Quelantes/farmacologia , Corpo Estriado/efeitos dos fármacos , Dopamina/metabolismo , Ácido Edético/farmacologia , Gadolínio/farmacologia , Masculino , Proteínas do Tecido Nervoso/metabolismo , Ligação Proteica/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Vesículas Sinápticas/química , Vesículas Sinápticas/metabolismo , Trítio , Tiramina/metabolismoRESUMO
MPP-production and uptake by dopaminergic terminals are critical steps in MPTP-induced Parkinson-like disorder. We reported evidence for a specific uptake of MPP by synaptic vesicles from mouse striatum. Its regional distribution suggests it as a marker of the dopamine vesicular carrier. We decided to further characterize such an MPP uptake. Tetrabenazine inhibits the dopamine uptake both in the striatum and in the cerebellum with similar Km values suggesting an identify of the vesicular carrier in these areas. On the contrary, 3H-MPP vesicular uptake had in the striatum a t1/2 of 60 sec, but was not detectable at any time in the cerebellum. Moreover, MPP inhibited the uptake of 3H-DA (Ki: 1.6 +/- 0.03 microM) and 3H-NE (Ki 2.6 +/- 0.01 microM) in the striatum but not in the cerebellum, even at molar concentration. These pharmacological data indicate that in nondopaminergic areas the monoamine carrier may be similar but not identical from that located in dopaminergic areas.
Assuntos
1-Metil-4-fenilpiridínio/metabolismo , Animais , Transporte Biológico , Biomarcadores , Cerebelo/metabolismo , Corpo Estriado/metabolismo , Dopamina/metabolismo , Hipocampo/metabolismo , Masculino , Camundongos , Norepinefrina/metabolismo , TrítioAssuntos
1-Metil-4-Fenil-1,2,3,6-Tetra-Hidropiridina/metabolismo , 1-Metil-4-fenilpiridínio/metabolismo , Corpo Estriado/metabolismo , Anfetamina/metabolismo , Animais , Ligação Competitiva/efeitos dos fármacos , Dopamina/metabolismo , Técnicas In Vitro , Melaninas/metabolismo , Membranas/metabolismo , Camundongos , Monoaminoxidase/metabolismo , Terminações Nervosas/efeitos dos fármacos , Terminações Nervosas/metabolismo , Tiramina/metabolismoAssuntos
Envelhecimento , Encéfalo/efeitos dos fármacos , Exposição Ambiental , Glicoproteínas de Membrana , Proteínas de Membrana Transportadoras , Neurotoxinas/toxicidade , Animais , Proteínas de Transporte/efeitos dos fármacos , Proteínas da Membrana Plasmática de Transporte de Dopamina , Humanos , Proteínas do Tecido Nervoso/efeitos dos fármacos , Vesículas Sinápticas/efeitos dos fármacos , TrítioRESUMO
The induction of the early gene c-fos was evaluated through Fos immunohistochemistry in areas belonging to the extended amygdala after acute administration of two antidepressants, citalopram and imipramine. Both citalopram and imipramine at the dose of 5 and 20 mg/kg, respectively, induced Fos-like immunoreactivity (FLI) in the central amygdaloid nucleus, lateral division of the bed nucleus of the stria terminalis (BSTL), and interstitial nucleus of the posterior limb of the anterior commissure (IPAC). The shell of the nucleus accumbens, which forms a continuum with the central extended amygdala, showed a decrease of FLI after administration of either citalopram or imipramine. The mechanism of action and the brain areas affected by antidepressants are still a matter of debate. By showing that the central extended amygdala is a common site of action for two different antidepressant types, these results provide new insight into the mechanism of action of antidepressants.
Assuntos
Inibidores da Captação Adrenérgica/farmacologia , Tonsila do Cerebelo/efeitos dos fármacos , Antidepressivos/farmacologia , Proteínas do Tecido Nervoso/biossíntese , Proteínas Proto-Oncogênicas c-fos/biossíntese , Inibidores Seletivos de Recaptação de Serotonina/farmacologia , Tonsila do Cerebelo/metabolismo , Animais , Citalopram/farmacologia , Imipramina/farmacologia , Imuno-Histoquímica , Masculino , Núcleo Accumbens/efeitos dos fármacos , Núcleo Accumbens/metabolismo , Ratos , Ratos Sprague-DawleyRESUMO
Dithiocarbamate compounds are widely used agricultural fungicides that display low acute toxicity in mammals and that may become neurotoxic after prolonged exposure. Mancozeb, among other dithiocarbamates tested, proved to be the most potent (Ki= 0.27 microM) at noncompetitively inhibiting the in vitro ATP-dependent uptake of [3H]glutamate in rat cortical vesicles. Furthermore, mancozeb partially (20%) inhibited the ATP-dependent uptake of [14C]methylamine, used as an index for the vesicular transmembrane proton gradient (DeltapH), and evoked its efflux from organelles previously incubated with the 3H-labeled marker. Meanwhile, the vesicular uptake of 36chloride- anions whose concentrations regulate the transmembrane potential gradient (DeltapsiSV) was not impaired. The dithiocarbamate effects on the vesicular transport of [3H]glutamate thus appeared to involve mainly the DeltapH gradient rather than the potential gradient. Dithiocarbamate metabolites, the potent neurotoxin carbon disulfide included, did not affect the uptake process, thus implying the relevance for inhibition of the persistence, if any, of parent compounds in the brain. The present novel and potent in vitro interferences of selected dithiocarbamate pesticides with the vesicular transport of glutamate, if representative of in vivo alterations, may play some role in the probably complex origin of dithiocarbamate neurotoxicity.
Assuntos
Ácido Glutâmico/metabolismo , Praguicidas/farmacologia , Vesículas Sinápticas/efeitos dos fármacos , Tiocarbamatos/farmacologia , Trifosfato de Adenosina/metabolismo , Animais , Transporte Biológico/efeitos dos fármacos , Encéfalo/efeitos dos fármacos , Encéfalo/metabolismo , Cloretos/metabolismo , Técnicas In Vitro , Masculino , Maneb/farmacologia , Metilaminas/metabolismo , Ratos , Ratos Sprague-Dawley , Ratos Wistar , Vesículas Sinápticas/metabolismo , Zineb/farmacologiaRESUMO
Because 1-methyl-4-phenyl-2,3-dihydropyridinium ion (MPP+) appears to damage the dopaminergic neuron and cause neuronal death, we characterized [3H]MPP+ binding sites in mouse brain membranes. Among several compounds tested, debrisoquin [3,4-dihydro-2(1H)-isoquinolinecarboxamidine] and some analogues were able to antagonize [3H]MPP+ binding. Debrisoquin is able to block adrenergic transmission and inhibit the activity of monoamine oxidase A (MAO-A). We found a certain correlation between the ability of these agents to displace [3H]MPP+ from its binding sites and their capacity to inhibit MAO-A activity. These data and the finding of a higher number of [3H]MPP+ binding sites in human placenta compared to mouse brain suggest that these sites may correspond to MAO-A enzymes. Recently it has been demonstrated in human brain that neurons in regions rich in catecholamines are positive for MAO-A. Accordingly, we suggest MAO-A as a possible accumulation site of MPP+ within the dopaminergic neuron. We also indicate the chemical structural requirement associated with the best binding of debrisoquin analogues with [3H]MPP+ sites. It would be reasonable to test the effects of debrisoquin-like drugs able to pass the blood-brain barrier on 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine toxicity.
Assuntos
Encéfalo/metabolismo , Compostos de Piridínio/metabolismo , Animais , Sítios de Ligação , Ligação Competitiva , Encéfalo/enzimologia , Fenômenos Químicos , Química , Debrisoquina/análogos & derivados , Feminino , Humanos , Íons , Masculino , Camundongos , Camundongos Endogâmicos , Monoaminoxidase/metabolismo , Placenta/metabolismo , TrítioRESUMO
The distribution and density of 3H-MPP+ binding sites were studied by in vitro quantitative autoradiography in the brain of the mouse, rat and monkey. The highest levels of 3H-MPP+ specific binding were observed in rat brain. The substantia nigra in rat and monkey, and the anterior caudate-putamen formation in mouse and monkey showed the lowest density of autoradiographic grains. The presence of a relatively high density of MPP+ sites in the hippocampus of all species studied could be of interest to explain some effects of MPTP administration on convulsions caused by chemoconvulsants. The finding of a 60-70% reduction of 3H-MPP+ binding sites in the rat caudate-putamen, on the side of quinolinic acid infusion and no changes after 6-hydroxydopamine lesion of dopaminergic nigrostriatal neurons suggests the presence of these sites mainly on striatal cells. The results suggest that the distribution of MPP+ binding sites in brain would not seem to be related to MPTP toxicity.
Assuntos
Química Encefálica , Callithrix/genética , Camundongos/metabolismo , Compostos de Piridínio/metabolismo , Ratos/metabolismo , 1-Metil-4-Fenil-1,2,3,6-Tetra-Hidropiridina/farmacocinética , Animais , Sítios de Ligação , Corpo Estriado/efeitos dos fármacos , Corpo Estriado/metabolismo , Feminino , Processamento de Imagem Assistida por Computador , Masculino , Monoaminoxidase/metabolismo , Oxidopamina/toxicidade , Compostos de Piridínio/farmacocinética , Ratos Endogâmicos/metabolismo , Especificidade da EspécieRESUMO
The effect of ritanserin on dopamine (DA) re-uptake and efflux was studied in rat frontal cortex synaptosomes. When compared to other 5HT2 receptor antagonists such as ketanserin and risperidone or DA D2 receptor antagonists such as haloperidol and raclopride, the effect of ritanserin proved to be more potent. Ritanserin blocked the DA transporter with a Ki of 0.18 +/- 0.06 microM, similar to cocaine (0.11 +/- 0.005 microM), while ketanserin had a Ki of 0.93 +/- 0.045; haloperidol of 2.07 +/- 0.12; risperidone of 18.01 +/- 0.62 and raclopride of 24.01 +/- 1.55. In addition, 15 min from its local application to the synaptosomes, ritanserin potently released [3]H-DA leaving only 29.6 +/- 1.6% of DA content, while ketanserin effect was equal to 46.5 +/- 0.9%; haloperidol to 70.4 +/- 2.2% and risperidone to 73.9 +/- 1.5%, all tested at the dose of 10 microM. Cocaine had no effect on DA efflux. These results suggest that ritanserin has a intrinsic dopaminergic effect which may help to explain its reported improvement on mood, cognition and negative symptoms of schizophrenia.
Assuntos
Dopamina/farmacocinética , Lobo Frontal/metabolismo , Receptores de Serotonina/metabolismo , Ritanserina/farmacologia , Antagonistas da Serotonina/farmacologia , Animais , Clozapina/farmacologia , Cocaína/farmacologia , Antagonistas de Dopamina/farmacologia , Inibidores da Captação de Dopamina/farmacologia , Lobo Frontal/efeitos dos fármacos , Haloperidol/farmacologia , Ketanserina/farmacologia , Masculino , Ratos , Ratos Sprague-Dawley , Risperidona/farmacologia , Sinaptossomos/efeitos dos fármacos , Sinaptossomos/metabolismo , TrítioRESUMO
Acute intoxication and chronic therapy with the alcohol consumption deterrent dithiocarbamate disulfiram have been associated with several neurological complications perhaps involving the impairment of neurotransmitter pathways. In this study we have tested the hypothesis that dopaminergic malfunction is a critical component in disulfiram-evoked neurotoxicity. Disulfiram antagonized the in vitro striatal binding of [3H]tyramine, a putative marker of the vesicular transporter for dopamine, and the uptake of [3H]dopamine into striatal synaptic vesicles, with inhibitory constants (Ki) in the range of reported blood dithiocarbamate levels in treated alcoholics. Furthermore, disulfiram provoked a loss of radioactivity from [3H]dopamine-preloaded striatal vesicles, when added directly to the incubation mixture. Several metal-containing fungicide analogs were also potent displacers of specifically bound [3H]tyramine. Diethyldithiocarbamate (DDC), the major metabolite of disulfiram, had none of these effects. The intraperitoneal injection of a high dose of disulfiram and DDC into rats, mimicking acute intoxication, induced in vivo overflow of striatal dopamine from both a reserpine-sensitive (vesicular) and an alpha-methyl-p-tyrosine-sensitive (cytoplasmic) pool. The vesicular component of in vivo dopamine release resulted mainly from a direct activity of disulfiram, on the organelles (interaction with the carrier for dopamine plus membrane permeabilization) and indirectly through the mediation of serotonergic 5-HT3 receptors. DDC acted poorly at the vesicle membrane, and the in vivo releasing effect of dopamine was only partially prevented by the inhibition of 5-HT3 receptors, thus suggesting the role of additional mechanisms. It is concluded that disulfiram intoxication may acutely disrupt dopamine balance, an effect probably underlying some of the central neurotoxic, extrapyramidal symptoms associated with dithiocarbamate overdose.
Assuntos
Dissuasores de Álcool/toxicidade , Antídotos/toxicidade , Corpo Estriado/efeitos dos fármacos , Dissulfiram/toxicidade , Ditiocarb/toxicidade , Dopamina/metabolismo , Ácido 3,4-Di-Hidroxifenilacético/metabolismo , Inibidores da Captação Adrenérgica/farmacologia , Dissuasores de Álcool/administração & dosagem , Animais , Antídotos/administração & dosagem , Ligação Competitiva , Bovinos , Permeabilidade da Membrana Celular/efeitos dos fármacos , Corpo Estriado/metabolismo , Citoplasma/efeitos dos fármacos , Citoplasma/metabolismo , Dissulfiram/administração & dosagem , Ditiocarb/administração & dosagem , Inibidores Enzimáticos/farmacologia , Injeções Intraperitoneais , Masculino , Metiltirosinas/farmacologia , Organelas/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Receptores de Serotonina/efeitos dos fármacos , Receptores de Serotonina/metabolismo , Receptores 5-HT3 de Serotonina , Reserpina/farmacologia , Vesículas Sinápticas/efeitos dos fármacos , Vesículas Sinápticas/metabolismo , Tiramina/metabolismo , alfa-MetiltirosinaRESUMO
Intoxication with the alcohol-aversive drug disulfiram (Antabuse) and related dithiocarbamates may provoke neuropathies and, in some cases, damage the basal ganglia. Rats received a single administration of disulfiram (7 and 500 mg kg-1 i.p.) and equimolar doses (4 and 290 mg kg-1 i.p.) of its metabolite diethyldithiocarbamate (DDC), roughly corresponding to the daily maximum dose in alcohol abusers or to an estimated nonlethal overdose, respectively. The striatal, extracellular levels of glutamate in freely moving rats previously implanted with a microdialysis probe increased after low and intoxicating doses of disulfiram (126 +/- 3% and 154 +/- 10% of basal values, respectively) and DDC as well (135 +/- 10% and 215 +/- 14%, respectively), a partially Ca++-dependent effect. The prolonged (>7 hr) disulfiram-induced increase in glutamate observed in vivo may reflect the in vitro disulfiram-evoked release of glutamate from striato-cortical synaptic vesicles, where the drug nonspecifically inhibited (Ki approximately 4 microM) the uptake function and abolished the transmembrane proton gradient (DeltapH). In contrast, DDC did not seem to affect DeltapH. The prompt DDC-provoked increase in extracellular levels of glutamate was prevented by 7-nitroindazole, an in vivo specific inhibitor of neuronal nitric oxide synthase, which suggests that the thiol metabolite also acts via the nitric oxide synthesis. At variance, the short-acting 7-nitroindazole did not prevent the sustained in vivo effects of disulfiram and of DDC putatively formed with time. These findings provide new evidence for differential mechanisms underlying disulfiram- and DDC-induced increases in striatal glutamate release. Present glutamatergic changes, although not appearing dramatic enough to represent the only cause for neuronal damage from disulfiram overdose, might contribute to the drug neurotoxicity.
Assuntos
Dissuasores de Álcool/intoxicação , Antídotos/intoxicação , Dissulfiram/intoxicação , Ácido Glutâmico/metabolismo , Animais , Transporte Biológico/efeitos dos fármacos , Ditiocarb/intoxicação , Masculino , Metilaminas/metabolismo , Ratos , Ratos Sprague-Dawley , Córtex Visual/efeitos dos fármacos , Córtex Visual/fisiologiaRESUMO
The effects of prenatal CO exposure (150 ppm from days 0 to 20 of pregnancy) on the postnatal development of hippocampal neuronal NO synthase (nNOS) and haem-oxygenase (HO-2) isoform activities in 15-, 30- and 90-d-old rats were investigated. Unlike HO-2, hippocampal nNOS activity increased from postnatal days 15-90 in controls. Prenatal CO produced a long-lasting decrease in either nNOS or HO-2. The results suggest that the altered developmental profile of hippocampal nNOS and HO-2 activities could be involved in cognitive deficits and long-term potentiation dysfunction exhibited by rats prenatally exposed to CO levels resulting in carboxyhaemoglobin (HbCO) levels equivalent to those observed in human cigarette smokers.